Effect of lentiviruses carrying enhanced green fluorescent protein injected into the scala media through a cochleostomy in rats

Am J Otolaryngol. 2013 Jul-Aug;34(4):301-7. doi: 10.1016/j.amjoto.2012.12.011. Epub 2013 Mar 5.

Abstract

Purpose: The purposes of the current study were to assess the feasibility of post-auricular microinjection of lentiviruses carrying enhanced green fluorescent protein (EGFP) into the scala media through cochleostomies in rats, determine the expression of viral gene in the cochlea, and record the post-operative changes in the number and auditory function of cochlear hair cells (HCs).

Methods: Healthy rats were randomly divided into two groups. The left ears of the animals in group I were injected with lentivirus carrying EGFP (n=10) via scala media lateral wall cochleostomies, and the left ears of the animals in group II were similarly injected with artificial endolymph (n=10). Prior to and 30 days post-injection, auditory function was assessed with click-auditory brainstem response (ABR) testing, EGFP expression was determined with cochlear frozen sections under fluorescence microscopy, and survival of HCs was estimated based on whole mount preparations.

Results: Thirty days after surgery, click-ABR testing revealed that there were significant differences in the auditory function, EGFP expression, and survival of HCs in the left ears before and after surgery in the same rats from each group. In group I, EGFP was noted in the strial marginal cells of the scala media, the organ of Corti, spiral nerves, and spiral ganglion cells.

Conclusion: Lentiviruses were successfully introduced into the scala media through cochleostomies in rats, and the EGFP reporter gene was efficiently expressed in the organ of Corti, spiral nerves, and spiral ganglion cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Survival
  • Cochlea / surgery*
  • Cochlear Duct / surgery*
  • Cochlear Duct / virology
  • Disease Models, Animal
  • Evoked Potentials, Auditory, Brain Stem / genetics
  • Evoked Potentials, Auditory, Brain Stem / physiology
  • Gene Expression Regulation, Viral
  • Genetic Therapy / methods*
  • Green Fluorescent Proteins / pharmacology*
  • Hair Cells, Auditory / physiology
  • Injections, Intralesional
  • Lentivirus / genetics*
  • Male
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley
  • Sensitivity and Specificity

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins